Polygonatum sibiricum polysaccharide hydrogel preparation of dihydromyricetin and preparation method of polygonatum sibiricum polysaccharide hydrogel preparation

Abstract

The invention discloses a polygonatum polysaccharide hydrogel preparation of dihydromyricetin. In the preparation process of the polygonatum polysaccharide hydrogel preparation of dihydromyricetin, polygonatum polysaccharide is used as an adhesive, food-grade low-molecular organic acid is used as a pH regulator and a preservative, and a food-grade calcium chloride ion cross-linking agent is used as a gel structure control agent. The polygonatum sibiricum polysaccharide hydrogel of dihydromyricetin prepared by the invention is a good sustained-release preparation, has better hydrophilic effect, gastrointestinal stability effect, permeability effect and sustained-release effect on dihydromyricetin, and enhances the bioavailability of dihydromyricetin. Meanwhile, the natural bioactive substance, namely, the dihydroxyenzyme, has the characteristics of low side effect and easiness in obtaining, so that the implementation of the invention is of great significance to the development of chronic disease targeted drugs and the prevention and treatment of chronic diseases in the future by the dihydroxyenzyme.

Description

Technical Field

[0001] The invention relates to the technical field of food and medicine homologous to medicine and pharmaceutical preparations, and in particular to a dihydromyricetin polygonatum polysaccharide hydrogel preparation and a preparation method thereof. Background Art

[0002] In recent years, with the improvement of living standards and changes in lifestyles, more and more people suffer from chronic diseases such as diabetes, hyperuricemia, hypertension, obesity, coronary heart disease and chronic bronchitis, which are on the rise year by year and are getting younger and younger. These diseases are mainly related to unreasonable lifestyles and eating habits. Adding functional ingredients to food and dietary supplements is a useful way to avoid and alleviate the occurrence of chronic diseases. Polyphenols, flavonoids, carotenoids, vitamins, probiotics and polyunsaturated fatty acids are commonly used functional ingredients in food for improving nutritional interventions for chronic diseases. These functional ingredients in food have the advantages of enhancing nutritional functions, improving food quality and alleviating diseases, but their own defects such as low solubility, bad odor, easy degradation, poor absorption and poor targeting greatly limit their functional applications. The delivery system is used to deliver functional ingredients in food and has been widely used in many fields such as food and pharmaceutical industries. In recent years, more attention has been paid to the use of proteins, polysaccharides and other polymers to formulate stable and effective carrier preparations for the delivery of bioactive compounds, such as nanoparticles, emulsions, liposomes, hydrogels and microencapsulation, which are some common colloidal delivery systems, which have greatly improved the stability of functional ingredients in food.

[0003] Dihydromyricetin is an extract of rattan tea (a wild woody vine of the genus Agave in the family Grapevine). Rattan tea was originally used as a daily drinking tea by the people. The Herbal Prescriptions states: "Rattan tea leaves are sweet and warm, can relieve thirst, relieve all kinds of bloating, relieve edema and relieve poison, and can be used as tea to relieve constipation and bowel movement". The main active ingredients of rattan tea are flavonoids, which have many unique effects such as scavenging free radicals, anti-oxidation, anti-thrombosis, anti-tumor, and anti-inflammatory. Dihydromyricetin is a relatively special flavonoid. In addition to the general characteristics of flavonoids, it also has the effects of relieving alcohol poisoning, preventing alcoholic liver disease, fatty liver disease, inhibiting liver cell deterioration, and reducing the incidence of liver cancer. Dihydromyricetin especially exhibits a broad spectrum of anti-tumor activity. When used in combination with tumor chemotherapy drugs, it can improve the efficacy and compliance of chemotherapy drugs by enhancing the inhibitory and killing effects of chemotherapy drugs on tumors and reducing the adverse reactions of chemotherapy drugs, providing a certain reference significance for the combined application of clinical anti-tumor drugs.

[0004] Polygonatum sibiricum is a plant material that is both a medicine and food. It is rich in starch, vitamins, mineral elements and a variety of active substances. Polygonatum sibiricum has been used as both a medicine and a food since ancient times. It has the effects of "relaxing the stomach and replenishing qi, harmonizing the five internal organs; filling muscles, strengthening the bone marrow, and doubling its strength; not aging for many years, and bright color; white hair becomes blacker, and teeth fall out and grow again". It is called "the surplus food of the immortals". Polygonatum sibiricum polysaccharide is the main functional component of Polygonatum sibiricum. Studies have shown that it has certain antioxidant, immunomodulatory, anti-fatigue, antiviral, blood sugar and blood lipid regulation, antibacterial and anti-inflammatory activities, and has a protective effect on various forms of oxidative damage and cell aging. In addition, Polygonatum sibiricum polysaccharide can adhere to the mucosal surface, and it has a tight connection between epithelial cells, so it prolongs the residence time in the intestine and enhances the potential of cell permeability. It has a good development and application prospect in functional food and medical care. Polygonatum sibiricum polysaccharide has multiple activities, and has good water solubility, stability and biological activity. It is an ideal food additive and can be used as a natural thickener and flavoring agent. It is prepared into a thermosensitive hydrogel to enable it to play a synergistic role in anti-tumor, hypoglycemic, immune regulation, etc., and reduce the dosage of the main drug.

[0005] Calcium chloride is an indispensable raw material in the food industry, and the daily intake per person is between 160 and 345 mg. It plays an important role in the food industry, mainly used as a stabilizer and coagulant, thickener, and nutritional enhancer. It has a wide range of applications, including but not limited to soy products, cream, soft drinks, jams, water preparation, and processing aids for the food industry. Food-grade calcium chloride can exist in various tissues of the human body in the form of salt, and has multiple physiological functions such as regulating the excitability of nerves, myocardium, and muscles, and participating in muscle contraction, cell secretion, and coagulation processes. In addition, it can also solidify soluble pectin into gel-like insoluble calcium pectinate, thereby maintaining the crispness and hardness of processed fruit and vegetable products. This property makes it an ideal gel structure controller. When dihydromyricetin is added to calcium-containing substances, since dihydromyricetin is slightly soluble in water, it can be added by emulsification with an emulsifier to improve its solubility. In addition, Polygonatum sibiricum polysaccharide as an adhesive usually has a low viscosity, so it is difficult to maintain stability during the production process. After adding calcium chloride, its calcium ions can combine with the anions present in the polysaccharide molecules and brass compounds to undergo a complex reaction to form a cross-linked structure, so that the polygonatum polysaccharide and dihydromyricetin stick together to form a gel, thereby improving its viscosity and physical properties such as mechanical strength and storage stability. Summary of the invention

[0006] The purpose of the present invention is to provide a dihydromyricetin-containing polygonatum polysaccharide hydrogel preparation and a preparation method thereof. The prepared dihydromyricetin-containing polygonatum polysaccharide hydrogel is a good sustained-release preparation and has better water solubility, intestinal fluid stability, and higher bioavailability.

[0007] In order to achieve the above object, the specific technical solution adopted by the present invention is as follows:

[0008] A dihydromyricetin-polygonatum polysaccharide hydrogel preparation, the preparation materials of which include: dihydromyricetin, polygonatum polysaccharide, food-grade low-molecular organic acid and calcium chloride ion crosslinking agent.

[0009] The method for preparing the dihydromyricetin-containing polygonatum polysaccharide hydrogel comprises the following steps:

[0010] 1) mixing and dissolving polygonatum polysaccharide with water, stirring, and standing to obtain a polygonatum polysaccharide aqueous solution;

[0011] 2) adding a food-grade low-molecular organic acid to the polygonatum polysaccharide aqueous solution prepared in step 1) to obtain an acidic polygonatum polysaccharide aqueous solution;

[0012] 3) adding dihydromyricetin to the acidic polygonatum polysaccharide aqueous solution in step 2), firstly performing ultrasonic dispersion, and then stirring the mixed solution to obtain a dihydromyricetin-polygonatum polysaccharide mixed solution;

[0013] 4) adding a food-grade calcium chloride ion crosslinking agent to the dihydromyricetin-polygonatum sibiricum polysaccharide mixed solution in step 3);

[0014] 5) The mixed solution prepared in step 4) is centrifuged to remove most of the water, and then subjected to vacuum freeze-drying to finally obtain a polysaccharide hydrogel freeze-dried powder preparation of dihydromyricetin.

[0015] In the step 1), the concentration of polygonatum polysaccharide is 1 mg / mL to 100 mg / mL, and the stirring temperature is 20°C to 100°C.

[0016] In the step 2), the pH of the acidic polygonatum polysaccharide aqueous solution is 3 to 6.5.

[0017] In the step 3), the mass ratio of dihydromyricetin to polygonatum polysaccharide is 1:1 to 100:1, the ultrasonic time is 1 min to 60 min, the stirring temperature is 20° C. to 80° C., and the stirring time is 10 min to 240 min.

[0018] The concentration of the calcium chloride ion crosslinking agent in the solution obtained in step 4) is 0.1 mg / mL to 0.5 mg / mL.

[0019] In the step 5), the centrifugal speed is 100 r / min to 10000 r / min, the vacuum freeze drying time is 1 h to 50 h, the temperature is -20°C to -80°C, and the vacuum degree is 1 to 50 Pa.

[0020] The food-grade low-molecular organic acid includes one or more of malic acid, citric acid, fumaric acid and succinic acid.

[0021] The dihydromyricetin polygonatum polysaccharide hydrogel preparation of the present invention is composed of dihydromyricetin, polygonatum polysaccharide, food-grade low-molecular organic acid and calcium chloride ion crosslinking agent. In the preparation process, polygonatum polysaccharide is used as a binder, food-grade low-molecular organic acid is used as a pH regulator and preservative, and food-grade calcium chloride ion crosslinking agent is used as a gel structure controller. The prepared dihydromyricetin polygonatum polysaccharide hydrogel is a good sustained-release preparation, has better hydrophilicity effect, gastrointestinal stability effect, permeability effect and sustained-release effect on dihydromyricetin, and enhances the bioavailability of dihydromyricetin. At the same time, the natural bioactive substance dihydromyricetin has the characteristics of low side effects and easy acquisition, and has important application significance for the prevention and treatment of chronic diseases in the future.

[0022] The invention adopts all food-grade raw materials, has good biocompatibility, strong controllability, and no toxic effect on the body. After encapsulating dihydromyricetin, the performance of the drug can be more significantly improved. Its targeting and step-by-step stimulation responsiveness can reduce the side effects of the drug on normal tissues and improve the utilization rate of the drug, which is of great significance for the development of targeted drugs in the future.

[0023] The hydrogel preparation prepared by the present invention successfully improves the oral bioavailability of drugs for treating chronic diseases.

[0024] The present invention has no complicated processing process, strong operability and wide application scenarios. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 . Comparison of the dissolution phenomenon of DMY-PSP solution and DMY suspension in implementation case 1. The solution after DMY-PSP dissolution is clear and transparent;

[0026] Figure 2 . Comparison of the stability of DMY-PSP and DMY monomer in simulated intestinal fluid environment in Example 1;

[0027] Figure 3 . A graph showing the relationship between DMY concentration in plasma and time after oral administration of DMY-PSP solution and DMY suspension to rats in Example 1;

[0028] Figure 4 . A graph showing changes in blood glucose levels in diabetic rats induced by streptozotocin solution after different treatments in Example 1;

[0029] Wherein: DMY-PSP represents the polygonatum polysaccharide hydrogel preparation of dihydromyricetin; DMY represents dihydromyricetin. DETAILED DESCRIPTION

[0030] Example 1

[0031] like Figure 1-4 As shown, a 10 mg / mL polysaccharide was mixed with water, dissolved, stirred, and allowed to stand to obtain a polysaccharide aqueous solution, and the stirring temperature was 50°C. Then, malic acid was added to the prepared polysaccharide aqueous solution until the pH was 4. Dihydromyricetin was added to the acidic polysaccharide aqueous solution, wherein the mass ratio of dihydromyricetin to polysaccharide was 2:1, and ultrasonic dispersion was first performed for 10 min, and then the mixed solution was stirred at a stirring temperature of 50°C and a stirring time of 60 min, and then a dihydromyricetin-polysaccharide mixed solution was obtained. A food-grade calcium chloride ion crosslinker was added to the dihydromyricetin-polysaccharide mixed solution to make the concentration of the calcium chloride ion crosslinker in the solution reach 0.1 mg / mL. The prepared mixed solution was centrifuged to remove most of the water, the centrifugal speed was 10000 r / min, and then vacuum freeze-dried for 24 hours, the freeze-drying temperature was -50°C, and the vacuum degree was 10 pa, and finally a polysaccharide hydrogel freeze-dried powder preparation of dihydromyricetin was obtained.

[0032] A stability test was conducted in an artificial intestinal fluid environment (simulated intestinal fluid pH = 6.8 prepared according to the United States Pharmacopoeia). When dihydromyricetin (DMY) was added to the artificial intestinal fluid environment, the DMY monomer was rapidly degraded in the intestinal fluid, and the retention rate of DMY after 6 hours was only 15.3%. When the dihydromyricetin-polysaccharide hydrogel preparation (DMY-PSP) prepared in Example 1 of the present invention was added to the intestinal fluid environment, it was found that it had good stability, and the retention rate within 6 hours could still be maintained at 94.1%.

[0033] 20 rats (200±20g) were randomly divided into two groups. They were fasted for 12h before the experiment and had free access to water. Each group of rats was gavaged with 1mL of DMY suspension and DMY-PSP solution prepared by the present invention (DMY dosage was 20mg / kg bwt). After administration, blood was taken from the tail vein for analysis at different time periods. Analysis of DMY concentration in plasma revealed that the bioavailability of DMY-PSP was 6.92 times higher than that of DMY.

[0034] 30 rats (200 ± 20g) were randomly divided into three groups, and rat diabetes model was induced with streptozotocin solution (50mg / kg). Blood glucose level was measured using a blood glucose meter. Rats that showed fasting blood glucose levels exceeding 16.7mM 7 days after administration were considered diabetic rats and used for the following studies. Rats fasted for 12h and had free drinking water. Before the experiment, BGL (blood glucose level) was measured, and blood samples were collected from the tail vein of each rat. Diabetic rats were randomly divided into 3 groups, 10 in each group. Three groups of diabetic rats were orally given free insulin with an insulin dose of 50IU / kg, and oral DMY-PSP (prepared by Example 1) loaded insulin preparations, and the last group of diabetic rats were subcutaneously injected with free insulin solution (insulin dose of 5IU / kg). Considering the effect of anesthetics on blood glucose levels, anesthetics were not used during this period. Blood glucose levels were detected at the scheduled time after administration. Plasma for insulin measurement was obtained by collecting 0.2 mL blood samples from the tail vein (collected at 0, 0.5, 1, 1.5, 2, 3, 4, 5, 6 and 8 h).

[0035] Subcutaneous injection of free insulin has a strong hypoglycemic effect. 1.5 hours after subcutaneous injection, blood sugar level can be reduced to 31% of the initial level. But it quickly rises after 2 hours and returns to 82% of the initial level after 4 hours. In addition, oral administration of free insulin solution (insulin dose of 50 IU / kg) did not show a significant blood sugar lowering effect, which indicates that oral free insulin is ineffective. Oral administration of DMY-PSP loaded insulin solution found that after 2 hours, it dropped to 50% of the initial level and could ensure a stable reduction in blood sugar level until 8 hours.

[0036] Example 2

[0037] Polygonatum sibiricum polysaccharide with a concentration of 20 mg / mL was mixed and dissolved with water, stirred, and allowed to stand to obtain a polygonatum sibiricum polysaccharide aqueous solution, and the stirring temperature was 60°C. Then citric acid was added to the prepared polygonatum sibiricum polysaccharide aqueous solution until the pH was 4.5. Dihydromyricetin was added to the acidic polygonatum sibiricum polysaccharide aqueous solution, wherein the mass ratio of dihydromyricetin to polygonatum sibiricum polysaccharide was 1:1, ultrasonic dispersion was first performed, the ultrasonic time was 30 minutes, and then the mixed solution was stirred at a stirring temperature of 45°C and a stirring time of 90 minutes, and then a dihydromyricetin-polygonatum sibiricum polysaccharide mixed solution was obtained. Food-grade calcium chloride ion crosslinker was added to the dihydromyricetin-polygonatum sibiricum polysaccharide mixed solution to make the concentration of calcium chloride ion crosslinker in the solution reach 0.1 mg / mL. The prepared mixed solution was centrifuged to remove most of the water, the centrifugal speed was 10000r / min, and then vacuum freeze-dried for 48 hours, the freeze-drying temperature was -70°C, and the vacuum degree was 35pa, and finally a polygonatum sibiricum polysaccharide hydrogel freeze-dried powder preparation of dihydromyricetin was obtained.

[0038] Example 3

[0039] Polygonatum sibiricum polysaccharide with a concentration of 20 mg / mL was mixed and dissolved with water, stirred, and allowed to stand to obtain a polygonatum sibiricum polysaccharide aqueous solution, and the stirring temperature was 80°C. Fumaric acid was then added to the prepared polygonatum sibiricum polysaccharide aqueous solution until the pH was 3.5. Dihydromyricetin was added to the acidic polygonatum sibiricum polysaccharide aqueous solution, wherein the mass ratio of dihydromyricetin to polygonatum sibiricum polysaccharide was 5:1, ultrasonic dispersion was first performed, the ultrasonic time was 45 minutes, and then the mixed solution was stirred at a stirring temperature of 70°C and a stirring time of 120 minutes, and then a dihydromyricetin-polygonatum sibiricum polysaccharide mixed solution was obtained. Food-grade calcium chloride ion crosslinker was added to the dihydromyricetin-polygonatum sibiricum polysaccharide mixed solution to make the concentration of calcium chloride ion crosslinker in the solution reach 0.2 mg / mL. The prepared mixed solution was centrifuged to remove most of the water, the centrifugal speed was 9000r / min, and then vacuum freeze-dried for 50 hours, the freeze-drying temperature was -50°C, and the vacuum degree was 30pa, and finally a polygonatum sibiricum polysaccharide hydrogel freeze-dried powder preparation of dihydromyricetin was obtained.

[0040] Although the present invention has been described in detail with reference to the aforementioned embodiments, it is still possible for those skilled in the art to modify the technical solutions described in the aforementioned embodiments, or to make equivalent substitutions for some of the technical features therein. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A dihydromyricetin-containing polygonatum polysaccharide hydrogel preparation, characterized in that: The preparation materials include: dihydromyricetin, polygonatum sibiricum polysaccharide, food-grade low-molecular organic acid and calcium chloride ion cross-linking agent.

2. The method for preparing dihydromyricetin-containing polygonatum polysaccharide hydrogel according to claim 1, characterized in that: The following steps are involved: 1) mixing and dissolving polygonatum polysaccharide with water, stirring, and standing to obtain a polygonatum polysaccharide aqueous solution; 2) adding a food-grade low-molecular organic acid to the polygonatum polysaccharide aqueous solution prepared in step 1) to obtain an acidic polygonatum polysaccharide aqueous solution; 3) adding dihydromyricetin to the acidic polygonatum polysaccharide aqueous solution in step 2), firstly performing ultrasonic dispersion, and then stirring the mixed solution to obtain a dihydromyricetin-polygonatum polysaccharide mixed solution; 4) adding a food-grade calcium chloride ion crosslinking agent to the dihydromyricetin-polygonatum sibiricum polysaccharide mixed solution in step 3); 5) The mixed solution prepared in step 4) is centrifuged to remove most of the water, and then subjected to vacuum freeze-drying to finally obtain a polysaccharide hydrogel freeze-dried powder preparation of dihydromyricetin.

3. The method for preparing dihydromyricetin-containing polygonatum polysaccharide hydrogel according to claim 2, characterized in that: In the step 1), the concentration of polygonatum polysaccharide is 1 mg / mL to 100 mg / mL, and the stirring temperature is 20°C to 100°C.

4. The method for preparing dihydromyricetin-containing polygonatum polysaccharide hydrogel according to claim 2, characterized in that: In the step 2), the pH of the acidic polygonatum polysaccharide aqueous solution is 3 to 6.

5.

5. The method for preparing dihydromyricetin-containing polygonatum polysaccharide hydrogel according to claim 2, characterized in that: In the step 3), the mass ratio of dihydromyricetin to polygonatum polysaccharide is 1:1 to 100:1, the ultrasonic time is 1 min to 60 min, the stirring temperature is 20° C. to 80° C., and the stirring time is 10 min to 240 min.

6. The method for preparing dihydromyricetin-containing polygonatum polysaccharide hydrogel according to claim 2, characterized in that: The concentration of the calcium chloride ion crosslinking agent in the solution obtained in step 4) is 0.1 mg / mL to 0.5 mg / mL.

7. The method for preparing dihydromyricetin-containing polygonatum polysaccharide hydrogel according to claim 2, characterized in that: In the step 5), the centrifugal speed is 100 r / min to 10000 r / min, the vacuum freeze drying time is 1 h to 50 h, the temperature is -20°C to -80°C, and the vacuum degree is 1 to 50 Pa.

8. The method for preparing dihydromyricetin-containing polygonatum polysaccharide hydrogel according to claim 2, characterized in that: The food-grade low-molecular organic acid includes one or more of malic acid, citric acid, fumaric acid and succinic acid.

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